Laboratory Evolution Reveals the Metabolic and Regulatory Basis of Ethylene Glycol Metabolism by Pseudomonas putida KT2440

Wing Jin Li, Lahiru N. Jayakody, Mary Ann Franden, Matthias Wehrmann, Tristan Daun, Bernhard Hauer, Lars M. Blank, Gregg T. Beckham, Janosch Klebensberger, Nick Wierckx

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76 Scopus Citations


Pollution from ethylene glycol, and plastics containing this monomer, represent a significant environmental problem. The investigation of its microbial metabolism therefore provides insights into the environmental fate of this pollutant and also enables its utilization as a carbon source for microbial biotechnology. Here, we reveal the genomic and metabolic basis of ethylene glycol metabolism in Pseudomonas putida KT2440. Although this strain cannot grow on ethylene glycol as sole carbon source, it can be used to generate growth-enhancing reducing equivalents upon co-feeding with acetate. Mutants that utilize ethylene glycol as sole carbon source were isolated through adaptive laboratory evolution. Genomic analysis of these mutants revealed a central role of the transcriptional regulator GclR, which represses the glyoxylate carboligase pathway as part of a larger metabolic context of purine and allantoin metabolism. Secondary mutations in a transcriptional regulator encoded by PP_2046 and a porin encoded by PP_2662 further improved growth on ethylene glycol in evolved strains, likely by balancing fluxes through the initial oxidations of ethylene glycol to glyoxylate. With this knowledge, we reverse engineered an ethylene glycol utilizing strain and thus revealed the metabolic and regulatory basis that are essential for efficient ethylene glycol metabolism in P. putida KT2440.

Original languageAmerican English
Pages (from-to)3669-3682
Number of pages14
JournalEnvironmental Microbiology
Issue number10
StatePublished - 1 Oct 2019

Bibliographical note

Publisher Copyright:
© 2019 Society for Applied Microbiology and John Wiley & Sons Ltd.

NREL Publication Number

  • NREL/JA-2A00-74156


  • adaptive laboratory evolution
  • ethylene glycol
  • genomics
  • glyoxylate
  • Pseudomonas putida
  • reverse engineering


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